Study on the tissue clearing process using different agents by Mueller matrix microscope.

In this paper, we monitor the in vitro tissue clearing process of mouse dorsal skin immersed into two types of agents using Mueller matrix microscope. By Mueller matrix polar decomposition, we can see that the major difference between polarization changes due to two kinds of agents is the opposite trend of phase retardance with clearing. For the insight of the connection between different agents with the microstructural and optical changes of cleared tissues, we establish various models to mimic the dynamic process of microphysical features of tissues with clearing time. The mechanisms considered include refractive index matching, collagen shrinkage, more orderly fibers and birefringence variation. We compare the experimental results with simulations based on a single mechanism model and a combined model, respectively, which confirms that an individual possible mechanism cannot explain the polarization phenomena due to clearing. Also by simulations of various clearing models involving two possible mechanisms, we can speculate that formamide and saturated sucrose as agents have respective impacts on tissue features and then cause different polarization changes with clearing. Specifically, collagen shrinkage plus birefringence reduction can better explain the tissue cleared by formamide, and refractive index match plus increased birefringence model is likely to be a proper description of tissue cleared by sucrose. Both simulations and experiments also validate the potential of Mueller matrix microscope as a good tool to understand the interaction between clearing agents and tissues.